Department of Molecular Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
EMBO J. 1986 Nov;5(11):2775-80. doi: 10.1002/j.1460-2075.1986.tb04567.x.
Mitochondria of S-type cytoplasmic male sterile maize contain two linear double-stranded DNA molecules, S1 and S2. Two open reading frames (ORF1 and ORF2) are present in S2 DNA. Fragments from ORF1 were inserted into plasmids to achieve expression in Escherichia coli. Cells transformed with recombinant plasmids produced mRNA which hybridized with ORF1 and corresponding polypeptides were synthesized by in vivo and in vitro systems. Antiserum against a lacZ/S2 fusion protein precipitated the anticipated polypeptides from transformed E. coli cells and was therefore used to detect homologous peptide sequences in protein preparations of mitochondria from different maize cytoplasms. The antiserum detected a protein of 125 000 M(r) present in mitochondria from male sterile B73S but absent from the fertile B73N cytoplasm.
S 型细胞质雄性不育玉米的线粒体含有两条线性双链 DNA 分子,S1 和 S2。S2 DNA 中存在两个开放阅读框(ORF1 和 ORF2)。ORF1 的片段被插入质粒中,以在大肠杆菌中实现表达。用重组质粒转化的细胞产生了与 ORF1 杂交的 mRNA,并且通过体内和体外系统合成了相应的多肽。针对 lacZ/S2 融合蛋白的抗血清从转化的大肠杆菌细胞中沉淀出预期的多肽,因此用于检测来自不同玉米细胞质的线粒体蛋白制剂中的同源肽序列。该抗血清检测到存在于雄性不育 B73S 线粒体中但不存在于可育 B73N 细胞质中的 125000M(r)的蛋白质。